OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST

Background: The hepatitis B test can be conducted using the real-time PCR method. In chronic cases, the test is performed by detecting one of the Hepatitis B Virus (HBV) specific genes, HBcAg. The real-time PCR method requires optimization to obtain optimal results. Purpose: This research aims to de...

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Main Authors: Tanti Yulia Dewi, Fusvita Merdekawati, Ai Djuminar, Yogi Khoirul Abror
Format: Article
Language:English
Published: Universitas Airlangga 2024-11-01
Series:Journal of Vocational Health Studies
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Online Access:https://e-journal.unair.ac.id/JVHS/article/view/47868
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author Tanti Yulia Dewi
Fusvita Merdekawati
Ai Djuminar
Yogi Khoirul Abror
author_facet Tanti Yulia Dewi
Fusvita Merdekawati
Ai Djuminar
Yogi Khoirul Abror
author_sort Tanti Yulia Dewi
collection DOAJ
description Background: The hepatitis B test can be conducted using the real-time PCR method. In chronic cases, the test is performed by detecting one of the Hepatitis B Virus (HBV) specific genes, HBcAg. The real-time PCR method requires optimization to obtain optimal results. Purpose: This research aims to determine the optimal temperature and time for denaturation in the hepatitis B test using the real-time PCR method. Method: The research method used was a quasi-experiment involving variations in temperature (94 °C, 95 °C, and 96 °C) and time (10 seconds, 15 seconds, and 20 seconds) for denaturation. Data processing resulted in static group comparisons based on 27 primary data from the Cycle Threshold (CT) value. Result: Variations in temperature conditions, specifically at 94 °C, 95 °C, and 96 °C, combined with a denaturation time of 10 seconds, yielded the mean CT values of 26.495, 26.355, and 26.003, respectively. When the temperature conditions were maintained at 94 °C, 95 °C, and 96 °C, with a denaturation time of 15 seconds, yielded the mean CT values of 25.962, 25.641, and 25.396. Similarly, under temperature conditions of 94 °C, 95 °C, and 96 °C with a denaturation time of 20 seconds, yielded the mean CT values of 26.544, 26.505, and 25.830 were obtained. Conclusion: Optimal results in this study are obtained through the acquisition of the smallest CT value, namely at temperature conditions of 96 °C and 15 seconds.
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issn 2580-7161
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publishDate 2024-11-01
publisher Universitas Airlangga
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spelling doaj-art-cbf1dec318b24499a10f54306b6727f72025-08-20T03:04:42ZengUniversitas AirlanggaJournal of Vocational Health Studies2580-71612580-717X2024-11-018213113610.20473/jvhs.V8.I2.2024.131-13645955OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TESTTanti Yulia Dewi0https://orcid.org/0009-0009-6009-0259Fusvita Merdekawati1https://orcid.org/0009-0000-9293-9610Ai Djuminar2https://orcid.org/0009-0007-4510-2333Yogi Khoirul Abror3https://orcid.org/0009-0002-9835-7595Department of Medical Laboratory Technology, Bandung Ministry of Health Polytechnic, BandungDepartment of Medical Laboratory Technology, Bandung Ministry of Health Polytechnic, BandungDepartment of Medical Laboratory Technology, Bandung Ministry of Health Polytechnic, BandungDepartment of Medical Laboratory Technology, Bandung Ministry of Health Polytechnic, BandungBackground: The hepatitis B test can be conducted using the real-time PCR method. In chronic cases, the test is performed by detecting one of the Hepatitis B Virus (HBV) specific genes, HBcAg. The real-time PCR method requires optimization to obtain optimal results. Purpose: This research aims to determine the optimal temperature and time for denaturation in the hepatitis B test using the real-time PCR method. Method: The research method used was a quasi-experiment involving variations in temperature (94 °C, 95 °C, and 96 °C) and time (10 seconds, 15 seconds, and 20 seconds) for denaturation. Data processing resulted in static group comparisons based on 27 primary data from the Cycle Threshold (CT) value. Result: Variations in temperature conditions, specifically at 94 °C, 95 °C, and 96 °C, combined with a denaturation time of 10 seconds, yielded the mean CT values of 26.495, 26.355, and 26.003, respectively. When the temperature conditions were maintained at 94 °C, 95 °C, and 96 °C, with a denaturation time of 15 seconds, yielded the mean CT values of 25.962, 25.641, and 25.396. Similarly, under temperature conditions of 94 °C, 95 °C, and 96 °C with a denaturation time of 20 seconds, yielded the mean CT values of 26.544, 26.505, and 25.830 were obtained. Conclusion: Optimal results in this study are obtained through the acquisition of the smallest CT value, namely at temperature conditions of 96 °C and 15 seconds.https://e-journal.unair.ac.id/JVHS/article/view/47868denaturationhbcag hepatitis bpcr real-time
spellingShingle Tanti Yulia Dewi
Fusvita Merdekawati
Ai Djuminar
Yogi Khoirul Abror
OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
Journal of Vocational Health Studies
denaturation
hbcag
hepatitis b
pcr
real-time
title OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
title_full OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
title_fullStr OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
title_full_unstemmed OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
title_short OPTIMIZATION OF DENATURATION TEMPERATURE AND TIME USING REAL-TIME PCR METHOD IN HEPATITIS B TEST
title_sort optimization of denaturation temperature and time using real time pcr method in hepatitis b test
topic denaturation
hbcag
hepatitis b
pcr
real-time
url https://e-journal.unair.ac.id/JVHS/article/view/47868
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AT fusvitamerdekawati optimizationofdenaturationtemperatureandtimeusingrealtimepcrmethodinhepatitisbtest
AT aidjuminar optimizationofdenaturationtemperatureandtimeusingrealtimepcrmethodinhepatitisbtest
AT yogikhoirulabror optimizationofdenaturationtemperatureandtimeusingrealtimepcrmethodinhepatitisbtest